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Iranian Journal of Basic Medical Sciences، جلد ۱۷، شماره ۸، صفحات ۵۶۶-۵۷۰
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عنوان انگلیسی A vector-based system for the differentiation of mouse embryonic stem cells toward germ-line cells
چکیده انگلیسی مقاله Objective(s):To culture thein vitro mouse embryonic stem cells (mESCs) and to direct their differentiation to germ-line cells; in present study we used a vector backbone containing the fusion construct Stra8-EGFP to select differentiated ES cells that entered meiosis. Retinoic acid was used to differentiate embryonic stem cells to germ cells. Materials and Methods: A fragment of Stra8 gene promoter (-1400 to +7) was inserted in ScaI/HindIII multiple cloning site of pEGFP-1 vector. Theelectroporationwas done on embryonic stem cells and positive colonies were selected as puromycin-resistant after three weeks of treatment with puromycin. All-trans retinoic acid (RA) was used for differentiation of mESCs at final concentration of 10-5M. The expression of protamine 1 (Prm1) gene was checked as post meiotic marker in differentiated mESCs after 5, 10, 15, 21 and 30 days after RA induction. Results: The PCR amplification by specific primers for Stra8-EGFP fusion gene was detected in DNA sample from mESCs after electroporation and puromycin treatment. GFP-positive mESC colonies were observed after 72 hr RA induction. The protamine 1 gene was expressed after 21 days of RA induction. Conclusion: In this study, we demonstrated the in vitro generation of mouse embryonic stem cells to germ cells by using a backbone vector containing the fusion gene Stra8-EGFP. The Stra8 gene is a retinoic acid-responsive protein and is able to regulate meiotic initiation.
کلیدواژه‌های انگلیسی مقاله
نویسندگان مقاله رضا ابراهیم زاده وصال | reza ebrahimzadeh vesal
department of medical genetics, faculty of medicine, tehran university of medical sciences, tehran, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی تهران (Tehran university of medical sciences)

محمد علی hokrgozar | mohammad ali hokrgozar
national cell bank of iran, pasteur institute of iran, tehran, iran

سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)

کریم nayernia | karim nayernia
institute of human genetics, north east england, stem cell institute, international center for life, newcastle university, newcastle, uk


لادن تیموری تولابی | ladan teimoori toolabi
molecular medicine department, biotechnology research center, pasteur institute of iran

سازمان اصلی تایید شده: انستیتو پاستور ایران (Pasteur institute of iran)

محمد میریونسی | mohammad miryounesi
genomic research center, shahid beheshti university of medical sciences, tehran, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی شهید بهشتی (Shahid beheshti university of medical sciences)

سیدمهدی nourashrafeddin | seyedmehdi nourashrafeddin
magee-womens research institute amp;amp; foundation, university of pittsburgh medical sciences, pittsburgh, pa 15213, usa


نجمه رنجی | najmeh ranji
department of genetics, faculty of sciences, islamic azad university, rasht branch, rasht, iran

سازمان اصلی تایید شده: دانشگاه آزاد اسلامی علوم و تحقیقات (Islamic azad university science and research branch)

محمد حسین مدرسی | mohammad hosein modarressi
department of medical genetics, faculty of medicine, tehran university of medical sciences, tehran, iran

سازمان اصلی تایید شده: دانشگاه علوم پزشکی تهران (Tehran university of medical sciences)

نشانی اینترنتی http://ijbms.mums.ac.ir/article_3190.html
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